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Team:NTNU Trondheim/Notebook
From 2014.igem.org
(Difference between revisions)
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</h6> | </h6> | ||
<div class="nb-tech">Transformation was done by heat-shock. A plasmid containing ampicillin resistance was used, and the transformed cells were incubated overnight on LB plates with ampicillin. Plates showed a bacterial blanket the next day; the cells were apparently super competent.</div> | <div class="nb-tech">Transformation was done by heat-shock. A plasmid containing ampicillin resistance was used, and the transformed cells were incubated overnight on LB plates with ampicillin. Plates showed a bacterial blanket the next day; the cells were apparently super competent.</div> | ||
| - | <p> Test of transformation efficiency of competent E. coli DH5α from June 6th. | + | <p> Test of transformation efficiency of competent <i>E. coli</i> DH5α from June 6th. |
</p> | </p> | ||
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</div> | </div> | ||
| + | <div class="entry nb-wet"> | ||
| + | <div> | ||
| + | <h6>June 11th | ||
| + | <div class="nb-onetech-i nohilite">show technical details</div> | ||
| + | </h6> | ||
| + | <div class="nb-tech">Only 1 µl was used for the transformation, the rest of the BioBrick stock solution was stored -20 °C. BBa_J23101 was located at plate 4, 17F. BBa_B0034 was located at plate 4, 1N and BBa_C0012 was located at plate 4, 1P. Plates showed decent growth the next day; transformation a success.</div> | ||
| + | <p> Rehydrated BioBrick BBa_J23101, BBa_B0034 and BBa_C0012, and transformed them into competent <i>E. coli</i> DH5α cells by heat-shock, and incubated the cultures on LB plates with ampicillin overnight. | ||
| + | |||
| + | </p> | ||
| + | </div> | ||
| + | </div> | ||
| + | |||
| + | <div class="entry nb-wet"> | ||
| + | <div> | ||
| + | <h6>June 12th | ||
| + | <div class="nb-onetech-i nohilite">show technical details</div> | ||
| + | </h6> | ||
| + | <div class="nb-tech">The SOC medium was clear the next day, meaning no growth. </div> | ||
| + | <p> Inoculated BioBrick colonies from June 11th in SOC medium containing ampicillin. | ||
| + | |||
| + | </p> | ||
| + | </div> | ||
| + | </div> | ||
| + | |||
| + | <div class="entry nb-wet"> | ||
| + | <div> | ||
| + | <h6>June 13th | ||
| + | <div class="nb-onetech-i nohilite">show technical details</div> | ||
| + | </h6> | ||
| + | <div class="nb-tech">The failed inoculation attempt on June 12th could indicate something wrong with the LB plates with ampicillin. Growth of non-transformed cells supported this. Most likely the ampicillin was aliquotted to the medium at an elevated temperature, causing denaturation of the antibiotic. </div> | ||
| + | <p> Negative control of non-transformed <i>E. coli</i> DH5α on LB plates with ampicillin. | ||
| + | |||
| + | </p> | ||
| + | </div> | ||
| + | </div> | ||
Revision as of 13:52, 6 August 2014
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Team:Cornell/notebook
From 2013.igem.org
Notebook
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Wet Lab
Dry Lab
Human Practices
Wiki
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Week 23
(02/06 - 08/06)
June 3rd
show technical details
testtesttesttest test
- Prepared SOC and yB solutions for future lab work.
- Sterilized material and solutions needed for future lab work.
June 4th
show technical details
{{{tech}}}
Made LB plates with ampicillin and ampicillin + kanamycin.
June 5th
show technical details
{{{tech}}}
Inoculated E. coli DH5α in SOC medium overnight.
June 6th
show technical details
OD of culture was 0.3160 after 100 minutes.
Made competent E. coli DH5α cells.
Week 24
(09/06 - 15/06)
June 10th
show technical details
Transformation was done by heat-shock. A plasmid containing ampicillin resistance was used, and the transformed cells were incubated overnight on LB plates with ampicillin. Plates showed a bacterial blanket the next day; the cells were apparently super competent.
Test of transformation efficiency of competent E. coli DH5α from June 6th.
June 11th
show technical details
Only 1 µl was used for the transformation, the rest of the BioBrick stock solution was stored -20 °C. BBa_J23101 was located at plate 4, 17F. BBa_B0034 was located at plate 4, 1N and BBa_C0012 was located at plate 4, 1P. Plates showed decent growth the next day; transformation a success.
Rehydrated BioBrick BBa_J23101, BBa_B0034 and BBa_C0012, and transformed them into competent E. coli DH5α cells by heat-shock, and incubated the cultures on LB plates with ampicillin overnight.
June 12th
show technical details
The SOC medium was clear the next day, meaning no growth.
Inoculated BioBrick colonies from June 11th in SOC medium containing ampicillin.
June 13th
show technical details
The failed inoculation attempt on June 12th could indicate something wrong with the LB plates with ampicillin. Growth of non-transformed cells supported this. Most likely the ampicillin was aliquotted to the medium at an elevated temperature, causing denaturation of the antibiotic.
Negative control of non-transformed E. coli DH5α on LB plates with ampicillin.
